Three phenylenediamine isomers (including ortho-, meta- and para-derivatives) were electrochemically polymerized to give polyphenylenediamine (PPD) films on platinized glassy carbon electrodes. Amperometric glucose sensors were developed by immobilizing glucose oxidase (GOx) into these polymer matrices during polymerization. Effects of the polymerization potential, polymerization charge, monomer concentration, GOx concentration and Pt deposition charge on the performance of the enzyme electrode to glucose were investigated. These resulting GC/Pt/PPD-GOx electrodes showed rapid electrochemical responses to hydrogen peroxide and glucose, and very good anti-interference ability to ascorbic acid. Correlation between the electroanalytical behaviors of the enzyme electrodes and the polymer structures was examined.